The American University in

Cairo
Laboratory Safety
Biological Hazards

Introduction
Biological hazards, are unlike chemicals, in
that they can reproduce.
Exposure to a small number of organisms
can create serious problems.
There are many different ways you can be
exposed to biological hazards when
working in the laboratory.
Types of agent, its life cycle, and how it can
be transmitted.

Recognition of Biological
Hazards

Biological Hazards
Biological agents are living things, or
substances produced by living things, that
can cause illness or disease in humans.
Biological agents include: Bacteria, viruses,
and fungi as well as larger organisms such
as parasites and plants.

Routes of Entry and

Transmission
Biological agents enter the body by
inhalation, by ingestion or by absorption
through the skin.
Inhalation is the most common route of entry.
Once inside the body, infectious agents can
multiply and be passed from one person to
another.
The route of transmission may be direct or
indirect.

Direct Transmission of Infectious

Agents
Direct transmission takes place when there is
physical contact between an infected and noninfected person.
It can also occur if there is a projection of
droplets into the mucous membranes of the
eye, nose or mouth. (cough or sneeze)
It might also occur if a person is injected or
punctured with an infected needle or other
sharp object which is infected.

Indirect Transmission
May occur when infectious agents attach
themselves to materials or objects such as
food and water or cooking and eating
utensils.
Transmitted by an insect.
Transmitted through the air and are
hazardous if inhaled.

Evaluating Biohazards

Biosafety Levels
There are four biosafety levels (BSLs), each
based on the degree of hazard, type of
agent, and modes of transmission.

Biosafety Levels (BSLs)

BSL-1:
No known or minimal potential hazard of
exposure to infectious agents.
Practices, safety equipment, and facilities
are appropriate for undergraduate and
secondary educational training and teaching
laboratories.

Biosafety level 2
Moderate potential hazard (low risk) of
exposure to infectious agents.
Practices, equipment, and facilities are
applicable to clinical, diagnostic, teaching
and other facilities in which work is done
with the broad spectrum of indigenous
moderate risk agents present in the
community and associated with human
disease of varying severity.

Biosafety level 3
Practices, safety equipment, and facilities are
applicable to clinical, diagnostic, teaching,
research, or production facilities in which
work is done with indigenous or exotic agents
with a potential for respiratory transmission,
and which may cause serious and potentially
lethal infection.
Moderate risk of exposure to agents that can
cause serious or potentially lethal disease.

Biosafety Level 4
High individual risk of exposure to dangerous
or exotic agents which cause life-threatening
disease.
Practices, safety equipment, and facilities are
applicable for work with dangerous and exotic
agents which pose a high individual risk of
life-threatening disease, which may be
transmitted via the aerosol route, and for
which there in no available vaccine or therapy.

Waste
1- Biohazardous Waste
Human or animal specimen cultures from
medical and pathological labs, cultures and
stocks of infectious agents from research labs.
Waste from production of bacteria, viruses or
the use of spores, discarded live and
attenuated vaccines, culture dishes and
contaminated devices used to transfer, and
mix cultures.

Biohazardous Waste
Human surgery specimens or tissues removed
at surgery.
Animal parts, tissues, fluids, or carcasses
suspected of being contaminated with
infectious agents.
Waste which contains recognizable fluid
blood, fluid blood products, containers, or
equipment containing fluid blood or blood
from animals, having been infected with
diseases that are highly communcable to
humans.

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Medical waste
Pharmaceutical waste
Sharps waste

Control
CONTAINMENT
It is a safe methods for managing infectious
agents in the laboratory environment where
they are being handled or maintained.
The purpose of containment is to reduce or
eliminate exposure of lab workers, other
persons, and the outside environment to
potentially hazardous agents.

Elements of Containment
1- Laboratory practice and techniques.
2- Safety Equipment (Primary Barriers).
3- Facility Design (Secondary Barriers).

Laboratory Practices and

Techniques
Persons working with infectious agents or
potentially infected materials must be aware
of potential hazards, and trained in the
practices and techniques required for
handling materials Safely.
Biosafety Manual for each lab.
Comptent person must direct lab activities.

Safety Equipment
Primary Barriers

Biological Safety Cabinets.

Enclosed containers (safety centrifuge cup)
PPE
(gloves, coats, gown, shoe covers, boots,
respirators, face shields, safety glasses and
goggles.)

Facility Design
Secondary Barriers
Separation of the laboratory work area from
public access.
Availability of decontamination facilities
(autoclave)
Handwashing facilities
Special Ventilation systems
Airlocks (double door entrance).

Biological Safety Cabinets

Biological safety cabinets (BSCs) are designed
to protect you from splashes and aerosols that
are contaminated with microbes, bacteria, and
viruses that can infect you.
There are three major classes of BSCs.
Class I BSCs: Protect you and the environment.
Class II BSCs: protect you and the
environment, and they also protect the materials
from contamination from outside.

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Class III BSCs:
Give the highest available protection to you,
the environment, and the material.

Class I BSC
This is a ventilated
cabinet for personnel
protection with an
unrecirculated inward
airflow away from the
operator.
This unit should be
fitted with HEPA filter
to protect the
environment from
discharged agents.

BSC I

BSC II
This is a ventilated
cabinet for personnel,
product and
environmental
protection which
provides inward air
flow and HEPAfiltered supply and
exhaust air.

BSC II
Class II cabinets may
be of use with low to
moderate risk
biological agents,
minute quantities of
toxic chemicals, and
trace quantities of
radionuclides.

BSC II

BSC III
A class III cabinet is a
totally enclosed ventilated
cabinet which is gas-tight,
and maintained under
negative air pressure.
The supply air is HEPAFiltered and the exhaust air
has two HEPA filters in
serious.
Work is performed in the
cabinet by the use of
attached rubber gloves.

BSC III

Guidelines for Working in

Biological Safety Cabinets
Make sure cabinet is tested and certified
before use.
Avoid exposure to cabinets UV light.
Turn air on and keep it running, allow air to
filter for few minutes before starting work.
Make sure everything needed for the
experiment is inside the cabinet.
Do not block cabinets air grilles with
material and equipment.

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Segregate contaminated items (I.e. clean
items to the left, contaminated items to the
right.) to avoid mix-ups.
Waste collection containers such as small
bio-hazardous waste bags, should be placed
inside cabinet to avoid bringing
contaminated items outside of cabinet, and
out into the lab.

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Avoid frequent and rapid arm movement in
and out of cabinet, to prevent disturbing
airflow and bringing contaminants out to
the lab.
Always keep spill absorbent towels and
decontaminating solution close by or in the
cabinet.
Prevent extended reaching, place materials
as close as possible.

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Perform your work at least 6 inches (15 cm)
into the BSC to maintain optimal air flow
containment for material and personal
protection.
Always assume a proper posture. Use only
adjustable chair or stool with built-in foot
and armrest.

Safe Laboratory Practices

Do not eat, drink or store food in the
laboratory. Store food in designated areas
only.
Do not apply cosmetics in the laboratory.
Never mouth pipette, always use
appropriate pipetting devices.
Remove contaminated clothing immediately
after it comes in contact with infectious
material.

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Always wear appropriate attire in the
laboratory. No matter how small is the task
at hand, always use assigned protective
clothing and equipment.
Always wash hands after any laboratory
activity, after removal of gloves or other
protective clothing, and immediately after
contact with infectious material.

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Manipulate all infectious materials carefully to
avoid spills and production of aerosols and
droplets. Aerosol generating procedures must be
performed in appropriate enclosures (Fume hoods,
BSC).
Decontaminate work surfaces before use, after use
and immediately after spills of infectious material.
Use a 70% ethanol to wipe cabinets work surfaces
down, prior to and after the completion of each
work session. Decontaminate with a 10% chlorine
bleach solution if necessary, especially after spills.

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Always use disposable glass or plastic ware. If nondisposable items must be used, make sure to disinfect
items before cleaning. (use a 10% chlorine bleach
solution)
Restrict the use of needles, scalpels, syringes, razor
blades and other sharps to those procedures for which
there are no other alternatives. If they must be used,
avoid self-inoculation; do not re-sheath or try to
remove used needles; dispose of whole assembly in a
leak-proof, and puncture-resistant container
immediately after use. Never leave sharps unattended
on any work surface.

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Discard all contaminated disposable items in
autoclave bags. All infectious waste
including normal cells, culture media and
microorganisms must be autoclaved before
disposal in the normal trash. (See infectious
waste disposal procedure).
Keep accurate inventories and inform
Environmental Health and Safety Office of
any infectious (biological hazardous)
material you acquire, including storage
location.

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Heat resistant gloves must be available and
used when unloading autoclave.